论文标题:细胞外基质金属蛋白酶诱导因子在糖尿病肾病中的表达和意义
The Study of Extracellular Matrix Metalloproteinase Inducer in Diabetic Nephropathy
论文作者
论文导师 陈平圣；邓红,论文学位 硕士,论文专业 病理学与病理生理学
论文单位 东南大学,点击次数 253,论文页数 49页File Size932K
2006-04-01论文网 http://www.lw23.com/lunwen_1186337/
diabetic nephropathy; human glomerular mesangial cells; extracellular matrix metalloproteinase inducer; matrix metalloproteinase-2; mechanism
研究目的1.建立糖尿病大鼠模型,观察糖尿病大鼠肾脏细胞外基质金属蛋白酶诱导因子(extracellular matrix metalloproteinase inducer,EMMPRIN)表达的变化,探讨糖尿病肾病发生、发展的机制。2.观察高糖对体外培养的人肾小球系膜细胞细胞外基质金属蛋白酶诱导因子表达的影响,探讨糖尿病肾病肾小球细胞外基质(extracellular matrix,ECM)积聚的机制。 研究方法1.将SD大鼠随机分成正常对照组和糖尿病模型组。腹腔注射链脲佐菌素(55mg·kg-1)诱导糖尿病大鼠模型,在0,4,8周末分批处死大鼠,用免疫组化检测EMMPRIN,MMP-2,CTGF和PDGF表达,图象分析仪半定量分析。2.系膜细胞培养:系膜细胞分别在对照组糖浓度(5mmol/L)、高糖浓度(25mmol/L)的条件下培养0h、12h、24h和48h,借助免疫细胞化学、RT-PCR等方法检测细胞外基质金属蛋白酶诱导因子和基质金属蛋白酶2表达情况。 研究结果1.正常大鼠EMMPRIN,MMP-2表达在大鼠肾小球系膜细胞、内皮细胞、包曼氏囊壁层上皮细胞、肾小管上皮细胞,造模后4周末和8周末时糖尿病大鼠EMMPRIN,MMP-2比正常组下降(P<0.05),糖尿病大鼠CTGF,PDGF表达比正常组升高(P<0.05)。2.系膜细胞培养时,高糖组在24h,48h可使培养的系膜细胞EMMPRIN mRNA和蛋白与正常组相比下降(P<0.05)、高糖组在48h MMP-2 mRNA和蛋白表达与正常组相比下降(P<0.05)。 研究结论1.糖尿病肾病时肾小球细胞外基质的积聚不仅是由于ECM合成增多,ECM降解减少也起重要作用。2.证实糖尿病时肾小球EMMPRIN表达水平降低,这可能是引起细胞外基质降解减少的原因之一。3.高糖使培养的系膜细胞内EMMPRIN,MMP-2表达降低,高糖有可能通过EMMPRIN调节MMP-2的表达,增加肾小球ECM的聚集,促进糖尿病肾病的进展。
Objectives 1.To observe the effects of high glucose concentration on the expression of EMMPRIN in kidneys and to explore the mechanisms of diabetic nephropathy by examining the expression of EMMPRIN. 2.To observe the effects of high ambient glucose concentration on the expression of EMMPRIN,MMP-2 in human glomerular mesangial cells in vitro and to explore further the mechanism of glomerular extracellular matrix accumulation in diabetic nephropathy. Methods 1.Rat diabetic model was induced by streptozotocin (55mg/kg),and the histology of kidneys was observed and the expression of EMMPRIN,MMP-2,CTGF,PDGF was examined by immunohistochemistry. 2.The mesangial cells were cultured in the high glucose media for 0h,12h,24h and 48h, respectively.The expression of EMMPRIN and MMP-2 mRNA was measured by RT-PCR and the protein expression of EMMPRIN and MMP-2 was observed by immunocytochemistry. Results 1.EMMPRIN and MMP-2 were mainly expressed in the mesangial cells,endothelial cells, podocytes, parietal epithelial cells of Bowman’s capsule,and tubular cells. The expression of EMMPRIN and MMP-2 was significantly weaken in the glomeruli of diabetic rats than that of the control animals(P<0.05). The expression of PDGF and CTGF was significantly enhenced in the glomeruli of diabetic rats than that of the control animal. 2.Compared with control groups, the expression of mRNA and protein expression of EMMPRIN and MMP-2 in the mesangial cells cultured in high glucose decreased(P<0.05). Conclusions 1.The increasing synthesis of many matrix components was not the only reason for the accumulation of glomerular extracellar matrix (ECM) in diabetic nephropathy.The decreasing degradation of matrix also plays a very important role in the accumulation of ECM. 2.These results demonstrated for the first time that EMMPRIN is downregulated in diabetic nephropathy.Decreased EMMPRIN may contribute to decreased degradation of the matrix. 3.High glucose down-regulates the mRNA and protein expression of EMMPRIN and MMP-2 in the mesangial cells.The decreased expression of EMMPRIN may alter the expression of MMPs in mesangial cell,thus influence the degradation of ECM in diabetic nephropathy.
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